表达恶性疟原虫裂殖子表面蛋白-1片段的减毒鼠伤寒杆菌的构建

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目的 检测恶性疟原虫裂殖子表面蛋白 1基因在减毒鼠伤寒直菌X40 6 4株中的表达。方法 将 7个克隆有恶性疟原虫裂殖子表面蛋白 1基因片段的表达质粒 pQEM用氯化钙法转化到修饰系统正常、限制系统缺陷的鼠伤寒杆菌LB5 0 0 0中 ,再通过专一性噬菌体P2 2 转导入腺苷酸环化酶基因、环腺苷酸受体基因双重突变的减毒鼠伤寒杆菌X40 6 4株中 ,用质粒酶切、Westernblot鉴定构建的重组菌株、测定重组菌株的表达。结果 成功地构建了 7个重组减毒鼠伤寒杆菌X40 6 4(pQEM )株 ,重组pQEM质粒在减毒鼠伤寒杆菌X40 6 4株中获得了表达。结论 X40 6 4(pQEM )菌能作为疟疾疫苗的候选疫苗菌株 Objective To detect the expression of Plasmodium falciparum merozoite surface protein 1 gene in attenuated Salmonella typhimurium strain X40 6 4. Methods Seven plasmids harboring Plasmodium falciparum merozoite surface protein 1 (pQEM) were transformed by calcium chloride into LB5 0 0 0, which modified the system and restricted the systemic defects. Bacteriophage P2 2 was transduced into the attenuated Salmonella typhimurium X40 6 4 strain with the adenylate cyclase gene and the cyclic adenosine monophosphate receptor double mutation. The recombinant strain was identified by Western blot using the plasmid and the recombinant strain expression. Results Seven recombinant attenuated Salmonella typhimurium X40 6 4 (pQEM) strains were successfully constructed and the recombinant plasmid pQEM was expressed in attenuated Salmonella typhimurium X40 6 4 strain. Conclusion X40 6 4 (pQEM) bacteria can be used as candidate vaccine strains for malaria vaccine
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